Use of bacteriophage endolysin EL188 and outer membrane permeabilizers against Pseudomonas aeruginosa
Use of bacteriophage endolysin EL188 and outer membrane permeabilizers against Pseudomonas aeruginosa
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DOI：
10.1111/j.1365-2672.2010.04931.x
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发表时间：
2011-03-01
影响因子：
4
通讯作者：
Lavigne, R.
中科院分区：
文献类型：
--
作者：
Briers, Y.;Walmagh, M.;Lavigne, R.
目的：选择和评价一种合适的外膜透析剂与高效溶血噬菌体内毒素EL188联合使用灭活(多重耐药)铜绿假单胞菌。方法和结果：我们测试了内溶酶EL188和几种外膜透透剂对3种选择的铜绿假单胞菌的作用。具有不同抗药性的铜绿假单胞菌。我们一方面用疏水探针N-苯基萘胺和肽聚糖结合域与绿色荧光蛋白的重组融合蛋白分析OM的通透性，另一方面用细胞裂解实验分析OM的通透性。抑菌实验表明，106Ps的孵育。在10 mmoL L-1乙二胺四乙酸二钠和50mgml1内毒素EL188存在下，3中心点01+/-0中心点17和4中心点27+/-0中心点11个对数单位在30min内呈菌株依赖性灭活。将EL188浓度增加到250mgml-1，可进一步提高对抗生素抗性最强的菌株Br667(4中心点07+/-0中心点09对数单位)的灭活效果。结论：乙二胺四乙酸二钠二水合物是EL188降低Ps的最佳组合。这项研究的意义和影响：这项体外研究表明，编码内毒素的噬菌体作为酶生物制剂的应用范围不能局限于革兰氏阳性病原体。
Aims:To select and evaluate an appropriate outer membrane (OM) permeabilizer to use in combination with the highly muralytic bacteriophage endolysin EL188 to inactivate (multi-resistant) Pseudomonas aeruginosa.Methods and Results:We tested the combination of endolysin EL188 and several OM permeabilizing compounds on three selected Ps. aeruginosa strains with varying antibiotic resistance. We analysed OM permeabilization using the hydrophobic probe N-phenylnaphtylamine and a recombinant fusion protein of a peptidoglycan binding domain and green fluorescent protein on the one hand and cell lysis assays on the other hand. Antibacterial assays showed that incubation of 106Ps. aeruginosa cells ml-1 in presence of 10 mmol l-1 ethylene diamine tetraacetic acid disodium salt dihydrate (EDTA) and 50 mu g ml-1 endolysin EL188 led to a strain-dependent inactivation between 3 center dot 01 +/- 0 center dot 17 and 4 center dot 27 +/- 0 center dot 11 log units in 30 min. Increasing the EL188 concentration to 250 mu g ml-1 further increased the inactivation of the most antibiotic resistant strain Br667 (4 center dot 07 +/- 0 center dot 09 log units).Conclusions:Ethylene diamine tetraacetic acid disodium salt dihydrate was selected as the most suitable component to combine with EL188 in order to reduce Ps. aeruginosa with up to 4 log units in a time interval of 30 min.Significance and Impact of the Study:This in vitro study demonstrates that the application range of bacteriophage encoded endolysins as 'enzybiotics' must not be limited to gram-positive pathogens.